Antisik1

Treated hPASMCs and frozen isolated sections were fixed in 4% paraformaldehyde for 30 min, blocked with 5% bovine serum albumin (BSA) for 2 h at room temperature, and then incubated overnight at 4 °C with the following primary antibodies: antiSIK1 (dilution 1:50; Proteintech), antiYAP (dilution 1:50, Santa Cruz), antiα-SMA (dilution 1:200, Proteintech), and antiKi67 (dilution 1:200, Abcam). After several washes in PBS, the cells and sections were incubated with the following appropriate directly conjugated fluorescent secondary antibodies: fluorescein isothiocyanate (FITC)-conjugated goat antimouse immunoglobulin G (IgG) (H + L) (dilution 1:200, Servicebio), Cy3-conjugated donkey antirabbit IgG (H + L) (dilution 1:200, Servicebio) for 2 h at room temperature, and diamidino-2-phenylindole (DAPI) (dilution 1:50; Beyotime) according to the manufacturer's instructions. Images were visualized by laser scanning confocal microscopy (LSM 800; Carl Zeiss, Germany).